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J Chromatogr A. 1996 Jun 7;736(1-2):291-302.

Large-scale amino-acid analysis for proteome studies.

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Macquarie University Centre for Analytical Biotechnology, Macquarie University, Sydney, NSW, Australia.


Amino-acid analysis is a relatively new method for identification of proteins separated by two-dimensional gel electrophoresis and blotted onto polyvinylidene difluoride (PVDF) membranes. This article describes modified amino-acid analysis methods for this purpose. Streamlined sample handling is a key feature of the process. To minimise sample manipulation, a single vial is used for hydrolysis and the protein hydrolysate on PVDF membrane is extracted by a one-step procedure. The hydrolysate should not be stored for long periods before analysis. Applications of the technique are presented to demonstrate the identification procedure. This approach is the most cost-effective and time-effective first step in mass protein screening for a large-scale proteome project.

[Indexed for MEDLINE]

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